Ceramic Electron Microscopy Grids for Cell Culturing and Multiscale Imaging

用于细胞培养和多尺度成像的陶瓷电子显微镜网格

• 批准号: 8200685
• 负责人: Oleg G. Polyakov
• 金额: $ 54.71万
• 依托单位: SYNKERA TECHNOLOGIES, INC.
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-06-01 至 2013-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8200685
• 关键词: Address; Architecture; Automation; Biological; Biological Sciences; Biophysics; Case Study; Cell Culture Techniques; Cells; Cellular Structures; Cellular biology; Ceramics; Colorado; Communities; Cryoelectron Microscopy; Development; Electron Microscopy; Evaluation; Film; Freezing; Generations; Generic Drugs; Goals; Image; Imaging technology; Joints; Laboratories; Mainstreaming; Methods; Microscopy; National Institute of General Medical Sciences; Optics; Performance; Phase; Preparation; Process; Protocols documentation; Publications; Research; Research Institute; Research Personnel; Sampling; Specimen; Structure; Techniques; Technology; Time; Universities; base; bioimaging; cellular imaging; comparative; cost; design; electron tomography; imaging modality; improved; light microscopy; meetings; molecular imaging; molecular scale; novel; prototype; scale up; single molecule; success; three-dimensional modeling

DESCRIPTION (provided by applicant): Improved supporting technologies for imaging of molecular and supramolecular structures within cells are needed to facilitate cell biology research, and are sought by the National Institute of General Medical Sciences (specifically, its Division of Cell Biology and Biophysics). Multiscale imaging, using cryo-electron tomography (cryo-ET) on supramolecular structures and single molecules, has proven in recent years to be a unique and invaluable method for highthroughput characterization of the dynamic 3D architecture of cells. Electron microscopy (EM) grids, used as substrates for supporting the biological and biomolecular specimens being imaged, are a critical component associated with this imaging method. New EM grid technology that decreases sample preparation cost and time, improves sample generation from culturing to freezing for cryo-ET, and increases imaging quality will allow researchers to more efficiently explore cellular architecture, at higher throughput. Synkera proposes a novel class of ceramic EM grids that feature an integrated thin support film that is highly compatible with cell culturing, light microscopy and cryo-ET. The grids will facilitate high-throughput, multiscale imaging of sub-cellular architecture and offer key advantages over state-of-the art products. The grids are also expected to be a competitive alternative in many other EM and culturing applications a 6-month Phase I project successfully demonstrated feasibility of the proposed grids architecture, as well as their potential in culturing, cryo-ET and multiscale imaging. Phase II aims to build on this success by further developing fabrication processes and ceramic EM grids designs, to fully realize the potential for multiscale imaging in functional prototypes. At least four academic partners will aide in this development process. The ultimate goal of the proposed project is a complete line of ceramic EM grids for a broad range of EM applications, from bioimaging to materials characterization. PUBLIC HEALTH RELEVANCE: The project addresses imaging of molecules and cells via cryo-electron tomography (cryo-ET). Specifically, the target application is multiscale imaging via optical microscopy and cryo-ET of cellular, supramolecular and single-molecule structures, for generating 3D models of sub-cellular architecture. The development of a novel class of ceramic-based electron microscopy grids that facilitate this multiscale imaging is proposed. The proposed technology will offer greater capability over state-of-the- art products and help further streamline multiscale cellular imaging by simplifying the specimen preparation process and yielding superior imaging performance.

描述（由申请人提供）：为了促进细胞生物学研究，需要改进细胞内分子和超分子结构成像的支持技术，这是美国国家普通医学科学研究所（特别是其细胞生物学和生物物理学部门）所寻求的。近年来，利用低温电子断层扫描（cryo-ET）对超分子结构和单分子进行多尺度成像，已被证明是一种独特而宝贵的方法，可用于高通量表征细胞的动态3D结构。电子显微镜（EM）网格用作支撑被成像的生物和生物分子标本的基底，是与该成像方法相关的关键组成部分。新的EM网格技术降低了样品制备成本和时间，改善了从培养到冷冻冷冻的样品生成，并提高了成像质量，这将使研究人员能够以更高的吞吐量更有效地探索细胞结构。Synkera提出了一种新型的陶瓷EM网格，具有集成的薄支撑膜，与细胞培养，光学显微镜和冷冻电镜高度兼容。网格将促进亚细胞结构的高通量、多尺度成像，并提供优于最先进产品的关键优势。网格也有望在许多其他EM和培养应用中成为有竞争力的替代方案。为期6个月的第一阶段项目成功地证明了所提出的网格架构的可行性，以及它们在培养、低温et和多尺度成像方面的潜力。第二阶段的目标是在这一成功的基础上，进一步开发制造工艺和陶瓷电磁网格设计，以充分实现功能原型中多尺度成像的潜力。至少有四个学术合作伙伴将协助这一发展过程。拟议项目的最终目标是为广泛的EM应用提供完整的陶瓷EM网格线，从生物成像到材料表征。